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        If you use plots from MultiQC in a publication or presentation, please cite:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

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        About MultiQC

        This report was generated using MultiQC, version 1.22.2

        You can see a YouTube video describing how to use MultiQC reports here: https://youtu.be/qPbIlO_KWN0

        For more information about MultiQC, including other videos and extensive documentation, please visit http://multiqc.info

        You can report bugs, suggest improvements and find the source code for MultiQC on GitHub: https://github.com/MultiQC/MultiQC

        MultiQC is published in Bioinformatics:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

        A modular tool to aggregate results from bioinformatics analyses across many samples into a single report.

        Report generated on 2024-07-13, 16:02 CEST based on data in: /crex/proj/naiss2024-23-424/analysis/outputs/FastQC


        General Statistics

        Showing 132/132 rows and 4/6 columns.
        Sample Name% Dups% GCMedian Read LengthM Seqs
        ALAN_SRR27370365_1P
        76.4%
        50%
        98bp
        38.7M
        ALAN_SRR27370365_1U
        17.2%
        50%
        90bp
        0.0M
        ALAN_SRR27370365_2P
        75.2%
        50%
        98bp
        38.7M
        ALAN_SRR27370365_2U
        0.0%
        0%
        0bp
        0.0M
        ALAN_SRR27370371_1P
        81.8%
        49%
        98bp
        76.5M
        ALAN_SRR27370371_1U
        18.5%
        48%
        90bp
        0.0M
        ALAN_SRR27370371_2P
        80.5%
        49%
        98bp
        76.5M
        ALAN_SRR27370371_2U
        0.0%
        0%
        0bp
        0.0M
        ALAN_SRR27370375_1P
        80.3%
        50%
        98bp
        62.3M
        ALAN_SRR27370375_1U
        14.4%
        49%
        90bp
        0.0M
        ALAN_SRR27370375_2P
        78.8%
        50%
        98bp
        62.3M
        ALAN_SRR27370375_2U
        0.0%
        0%
        0bp
        0.0M
        ALAN_SRR27370376_1P
        77.9%
        49%
        98bp
        44.5M
        ALAN_SRR27370376_1U
        18.0%
        48%
        90bp
        0.0M
        ALAN_SRR27370376_2P
        76.3%
        49%
        98bp
        44.5M
        ALAN_SRR27370376_2U
        0.0%
        0%
        0bp
        0.0M
        ALAN_SRR27370382_1P
        80.0%
        50%
        98bp
        77.9M
        ALAN_SRR27370382_1U
        16.0%
        48%
        91bp
        0.0M
        ALAN_SRR27370382_2P
        78.2%
        50%
        98bp
        77.9M
        ALAN_SRR27370382_2U
        0.0%
        0%
        0bp
        0.0M
        SRR27370365_1
        76.4%
        50%
        98bp
        38.8M
        SRR27370365_2
        75.2%
        50%
        98bp
        38.8M
        SRR27370366_1
        79.5%
        49%
        98bp
        61.2M
        SRR27370366_2
        77.7%
        50%
        98bp
        61.2M
        SRR27370367_1
        80.9%
        50%
        98bp
        63.4M
        SRR27370367_2
        79.5%
        51%
        98bp
        63.4M
        SRR27370368_1
        77.9%
        49%
        98bp
        55.0M
        SRR27370368_2
        76.3%
        50%
        98bp
        55.0M
        SRR27370369_1
        78.1%
        49%
        98bp
        58.9M
        SRR27370369_2
        76.0%
        49%
        98bp
        58.9M
        SRR27370370_1
        81.0%
        50%
        98bp
        58.7M
        SRR27370370_2
        79.8%
        50%
        98bp
        58.7M
        SRR27370371_1
        81.8%
        49%
        98bp
        76.5M
        SRR27370371_2
        80.5%
        49%
        98bp
        76.5M
        SRR27370372_1
        84.1%
        49%
        98bp
        99.1M
        SRR27370372_2
        82.6%
        49%
        98bp
        99.1M
        SRR27370373_1
        81.0%
        48%
        98bp
        52.4M
        SRR27370373_2
        78.5%
        49%
        98bp
        52.4M
        SRR27370374_1
        78.5%
        50%
        98bp
        52.3M
        SRR27370374_2
        77.2%
        51%
        98bp
        52.3M
        SRR27370375_1
        80.3%
        50%
        98bp
        62.3M
        SRR27370375_2
        78.8%
        50%
        98bp
        62.3M
        SRR27370376_1
        77.9%
        49%
        98bp
        44.5M
        SRR27370376_2
        76.3%
        49%
        98bp
        44.5M
        SRR27370377_1
        77.8%
        49%
        98bp
        48.3M
        SRR27370377_2
        75.9%
        50%
        98bp
        48.3M
        SRR27370378_1
        79.9%
        49%
        98bp
        57.4M
        SRR27370378_2
        78.1%
        50%
        98bp
        57.4M
        SRR27370379_1
        80.8%
        50%
        98bp
        71.3M
        SRR27370379_2
        79.0%
        50%
        98bp
        71.3M
        SRR27370380_1
        80.5%
        49%
        98bp
        64.6M
        SRR27370380_2
        78.7%
        50%
        98bp
        64.6M
        SRR27370381_1
        80.4%
        49%
        98bp
        69.1M
        SRR27370381_2
        78.3%
        49%
        98bp
        69.1M
        SRR27370382_1
        80.0%
        50%
        98bp
        77.9M
        SRR27370382_2
        78.2%
        50%
        98bp
        77.9M
        SRR27370383_1
        76.1%
        50%
        98bp
        49.4M
        SRR27370383_2
        74.5%
        50%
        98bp
        49.4M
        SRR27370384_1
        78.0%
        49%
        98bp
        56.7M
        SRR27370384_2
        76.2%
        50%
        98bp
        56.7M
        SRR27370385_1
        79.1%
        50%
        98bp
        56.8M
        SRR27370385_2
        77.6%
        50%
        98bp
        56.8M
        SRR27370386_1
        77.7%
        50%
        98bp
        49.8M
        SRR27370386_2
        76.1%
        50%
        98bp
        49.8M
        control_SRR27370367_1P
        80.9%
        50%
        98bp
        63.3M
        control_SRR27370367_1U
        22.6%
        51%
        91bp
        0.0M
        control_SRR27370367_2P
        79.5%
        51%
        98bp
        63.3M
        control_SRR27370367_2U
        0.0%
        0%
        0bp
        0.0M
        control_SRR27370378_1P
        79.9%
        49%
        98bp
        57.4M
        control_SRR27370378_1U
        21.0%
        49%
        90bp
        0.0M
        control_SRR27370378_2P
        78.1%
        50%
        98bp
        57.4M
        control_SRR27370378_2U
        0.0%
        0%
        0bp
        0.0M
        control_SRR27370379_1P
        80.8%
        50%
        98bp
        71.2M
        control_SRR27370379_1U
        16.4%
        49%
        91bp
        0.0M
        control_SRR27370379_2P
        79.0%
        50%
        98bp
        71.2M
        control_SRR27370379_2U
        0.0%
        0%
        0bp
        0.0M
        control_SRR27370384_1P
        78.0%
        49%
        98bp
        56.7M
        control_SRR27370384_1U
        14.6%
        48%
        91bp
        0.0M
        control_SRR27370384_2P
        76.2%
        50%
        98bp
        56.7M
        control_SRR27370384_2U
        0.0%
        0%
        0bp
        0.0M
        control_SRR27370385_1P
        79.1%
        50%
        98bp
        56.8M
        control_SRR27370385_1U
        17.8%
        49%
        90bp
        0.0M
        control_SRR27370385_2P
        77.6%
        50%
        98bp
        56.8M
        control_SRR27370385_2U
        0.0%
        0%
        0bp
        0.0M
        noise_SRR27370366_1P
        79.5%
        49%
        98bp
        61.2M
        noise_SRR27370366_1U
        15.3%
        48%
        90bp
        0.0M
        noise_SRR27370366_2P
        77.7%
        50%
        98bp
        61.2M
        noise_SRR27370366_2U
        0.0%
        0%
        0bp
        0.0M
        noise_SRR27370370_1P
        81.0%
        50%
        98bp
        58.6M
        noise_SRR27370370_1U
        17.3%
        50%
        90bp
        0.0M
        noise_SRR27370370_2P
        79.8%
        50%
        98bp
        58.6M
        noise_SRR27370370_2U
        0.0%
        0%
        0bp
        0.0M
        noise_SRR27370372_1P
        84.1%
        49%
        98bp
        99.1M
        noise_SRR27370372_1U
        15.7%
        49%
        90bp
        0.0M
        noise_SRR27370372_2P
        82.6%
        49%
        98bp
        99.1M
        noise_SRR27370372_2U
        0.0%
        0%
        0bp
        0.0M
        noise_SRR27370373_1P
        81.0%
        48%
        98bp
        52.4M
        noise_SRR27370373_1U
        8.9%
        47%
        90bp
        0.0M
        noise_SRR27370373_2P
        78.5%
        49%
        98bp
        52.4M
        noise_SRR27370373_2U
        0.0%
        0%
        0bp
        0.0M
        noise_SRR27370374_1P
        78.5%
        50%
        98bp
        52.3M
        noise_SRR27370374_1U
        17.5%
        50%
        90bp
        0.0M
        noise_SRR27370374_2P
        77.2%
        51%
        98bp
        52.3M
        noise_SRR27370374_2U
        0.0%
        0%
        0bp
        0.0M
        noise_SRR27370381_1P
        80.4%
        49%
        98bp
        69.1M
        noise_SRR27370381_1U
        15.8%
        48%
        91bp
        0.0M
        noise_SRR27370381_2P
        78.3%
        49%
        98bp
        69.1M
        noise_SRR27370381_2U
        0.0%
        0%
        0bp
        0.0M
        soot_SRR27370368_1P
        77.9%
        49%
        98bp
        55.0M
        soot_SRR27370368_1U
        18.2%
        49%
        90bp
        0.0M
        soot_SRR27370368_2P
        76.3%
        50%
        98bp
        55.0M
        soot_SRR27370368_2U
        0.0%
        0%
        0bp
        0.0M
        soot_SRR27370369_1P
        78.1%
        49%
        98bp
        58.9M
        soot_SRR27370369_1U
        18.5%
        48%
        90bp
        0.0M
        soot_SRR27370369_2P
        76.0%
        49%
        98bp
        58.9M
        soot_SRR27370369_2U
        0.0%
        0%
        0bp
        0.0M
        soot_SRR27370377_1P
        77.8%
        49%
        98bp
        48.3M
        soot_SRR27370377_1U
        13.7%
        48%
        90bp
        0.0M
        soot_SRR27370377_2P
        75.9%
        50%
        98bp
        48.3M
        soot_SRR27370377_2U
        0.0%
        0%
        0bp
        0.0M
        soot_SRR27370380_1P
        80.5%
        49%
        98bp
        64.6M
        soot_SRR27370380_1U
        15.8%
        49%
        91bp
        0.0M
        soot_SRR27370380_2P
        78.7%
        50%
        98bp
        64.6M
        soot_SRR27370380_2U
        0.0%
        0%
        0bp
        0.0M
        soot_SRR27370383_1P
        76.1%
        50%
        98bp
        49.4M
        soot_SRR27370383_1U
        11.7%
        48%
        90bp
        0.0M
        soot_SRR27370383_2P
        74.5%
        50%
        98bp
        49.4M
        soot_SRR27370383_2U
        0.0%
        0%
        0bp
        0.0M
        soot_SRR27370386_1P
        77.7%
        50%
        98bp
        49.8M
        soot_SRR27370386_1U
        20.2%
        51%
        90bp
        0.0M
        soot_SRR27370386_2P
        76.1%
        50%
        98bp
        49.8M
        soot_SRR27370386_2U
        0.0%
        0%
        0bp
        0.0M

        FastQC

        Version: 0.11.9

        FastQC is a quality control tool for high throughput sequence data, written by Simon Andrews at the Babraham Institute in Cambridge.

        Sequence Counts

        Sequence counts for each sample. Duplicate read counts are an estimate only.

        This plot show the total number of reads, broken down into unique and duplicate if possible (only more recent versions of FastQC give duplicate info).

        You can read more about duplicate calculation in the FastQC documentation. A small part has been copied here for convenience:

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Quality Histograms

        The mean quality value across each base position in the read.

        To enable multiple samples to be plotted on the same graph, only the mean quality scores are plotted (unlike the box plots seen in FastQC reports).

        Taken from the FastQC help:

        The y-axis on the graph shows the quality scores. The higher the score, the better the base call. The background of the graph divides the y axis into very good quality calls (green), calls of reasonable quality (orange), and calls of poor quality (red). The quality of calls on most platforms will degrade as the run progresses, so it is common to see base calls falling into the orange area towards the end of a read.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Sequence Quality Scores

        The number of reads with average quality scores. Shows if a subset of reads has poor quality.

        From the FastQC help:

        The per sequence quality score report allows you to see if a subset of your sequences have universally low quality values. It is often the case that a subset of sequences will have universally poor quality, however these should represent only a small percentage of the total sequences.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Base Sequence Content

        The proportion of each base position for which each of the four normal DNA bases has been called.

        To enable multiple samples to be shown in a single plot, the base composition data is shown as a heatmap. The colours represent the balance between the four bases: an even distribution should give an even muddy brown colour. Hover over the plot to see the percentage of the four bases under the cursor.

        To see the data as a line plot, as in the original FastQC graph, click on a sample track.

        From the FastQC help:

        Per Base Sequence Content plots out the proportion of each base position in a file for which each of the four normal DNA bases has been called.

        In a random library you would expect that there would be little to no difference between the different bases of a sequence run, so the lines in this plot should run parallel with each other. The relative amount of each base should reflect the overall amount of these bases in your genome, but in any case they should not be hugely imbalanced from each other.

        It's worth noting that some types of library will always produce biased sequence composition, normally at the start of the read. Libraries produced by priming using random hexamers (including nearly all RNA-Seq libraries) and those which were fragmented using transposases inherit an intrinsic bias in the positions at which reads start. This bias does not concern an absolute sequence, but instead provides enrichement of a number of different K-mers at the 5' end of the reads. Whilst this is a true technical bias, it isn't something which can be corrected by trimming and in most cases doesn't seem to adversely affect the downstream analysis.

        Click a sample row to see a line plot for that dataset.
        Rollover for sample name
        Position: -
        %T: -
        %C: -
        %A: -
        %G: -

        Per Sequence GC Content

        The average GC content of reads. Normal random library typically have a roughly normal distribution of GC content.

        From the FastQC help:

        This module measures the GC content across the whole length of each sequence in a file and compares it to a modelled normal distribution of GC content.

        In a normal random library you would expect to see a roughly normal distribution of GC content where the central peak corresponds to the overall GC content of the underlying genome. Since we don't know the the GC content of the genome the modal GC content is calculated from the observed data and used to build a reference distribution.

        An unusually shaped distribution could indicate a contaminated library or some other kinds of biased subset. A normal distribution which is shifted indicates some systematic bias which is independent of base position. If there is a systematic bias which creates a shifted normal distribution then this won't be flagged as an error by the module since it doesn't know what your genome's GC content should be.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Base N Content

        The percentage of base calls at each position for which an N was called.

        From the FastQC help:

        If a sequencer is unable to make a base call with sufficient confidence then it will normally substitute an N rather than a conventional base call. This graph shows the percentage of base calls at each position for which an N was called.

        It's not unusual to see a very low proportion of Ns appearing in a sequence, especially nearer the end of a sequence. However, if this proportion rises above a few percent it suggests that the analysis pipeline was unable to interpret the data well enough to make valid base calls.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Length Distribution

        The distribution of fragment sizes (read lengths) found. See the FastQC help

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Duplication Levels

        The relative level of duplication found for every sequence.

        From the FastQC Help:

        In a diverse library most sequences will occur only once in the final set. A low level of duplication may indicate a very high level of coverage of the target sequence, but a high level of duplication is more likely to indicate some kind of enrichment bias (eg PCR over amplification). This graph shows the degree of duplication for every sequence in a library: the relative number of sequences with different degrees of duplication.

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        In a properly diverse library most sequences should fall into the far left of the plot in both the red and blue lines. A general level of enrichment, indicating broad oversequencing in the library will tend to flatten the lines, lowering the low end and generally raising other categories. More specific enrichments of subsets, or the presence of low complexity contaminants will tend to produce spikes towards the right of the plot.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Overrepresented sequences by sample

        The total amount of overrepresented sequences found in each library.

        FastQC calculates and lists overrepresented sequences in FastQ files. It would not be possible to show this for all samples in a MultiQC report, so instead this plot shows the number of sequences categorized as overrepresented.

        Sometimes, a single sequence may account for a large number of reads in a dataset. To show this, the bars are split into two: the first shows the overrepresented reads that come from the single most common sequence. The second shows the total count from all remaining overrepresented sequences.

        From the FastQC Help:

        A normal high-throughput library will contain a diverse set of sequences, with no individual sequence making up a tiny fraction of the whole. Finding that a single sequence is very overrepresented in the set either means that it is highly biologically significant, or indicates that the library is contaminated, or not as diverse as you expected.

        FastQC lists all the sequences which make up more than 0.1% of the total. To conserve memory only sequences which appear in the first 100,000 sequences are tracked to the end of the file. It is therefore possible that a sequence which is overrepresented but doesn't appear at the start of the file for some reason could be missed by this module.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Top overrepresented sequences

        Top overrepresented sequences across all samples. The table shows 20 most overrepresented sequences across all samples, ranked by the number of samples they occur in.

        Showing 20/20 rows and 3/3 columns.
        Overrepresented sequenceSamplesOccurrences% of all reads
        AATTTTCTTTAGTTGTTCTTCTGTCATCTGGGGCTTGCGCTTGACGAGGT
        16
        245
        0.0000%
        CAAAGCTGAGAGCGACGCCGGGGTGGGGAGGTGGGGAGGCAATTCTAGGG
        13
        180
        0.0000%
        CAACATCTGGCTTCTTGCAGCACTTTTCCATCATGGCAGTGAAACCCTCA
        20
        451
        0.0000%
        CAGATGTTCCTGTGCTCCTGGGGGCCTCCACCCCCATGGGAAAGGTTCCC
        22
        952733
        0.0180%
        CCATGTCCCCGTCACAGCACTCTTTGTGGACATCCTTCACTTTTTGGACC
        16
        191
        0.0000%
        CCTTCACCTCCTCCACGTCCTTGGTCAGCTCGGCGCGCAGGGACTCGGTG
        14
        172
        0.0000%
        CTCCACCAACTAAGAACGGCCATGCACCACCACCCACGGAATCGAGAAAG
        18
        297
        0.0000%
        CTGCTTCGCGCTCAGCCGGAGGAGCCTTGCACATTTTCTCATCAAAATCG
        21
        653
        0.0000%
        GAGAGCGACGCCGGGGTGGGGAGGTGGGGAGGCAATTCTAGGGGAAGAGG
        13
        195
        0.0000%
        GAGGCAAAGCTGAGAGCGACGCCGGGGTGGGGAGGTGGGGAGGCAATTCT
        16
        203
        0.0000%
        GGAGGCAAAGCTGAGAGCGACGCCGGGGTGGGGAGGTGGGGAGGCAATTC
        17
        311
        0.0000%
        GGCTTGCGCTTGACGAGGTTGACGAGCAATTTCAGCTCCCCTGCTTCGCG
        17
        234
        0.0000%
        GTAATTTTCTTTAGTTGTTCTTCTGTCATCTGGGGCTTGCGCTTGACGAG
        13
        175
        0.0000%
        GTCATCTGGGGCTTGCGCTTGACGAGGTTGACGAGCAATTTCAGCTCCCCTGCTTCGCGCTCAGCCGGAGGAGCC
        14
        176
        0.0000%
        TCCACCAACTAAGAACGGCCATGCACCACCACCCACGGAATCGAGAAAGA
        22
        1236
        0.0000%
        TCTGTCATCTGGGGCTTGCGCTTGACGAGGTTGACGAGCAATTTCAGCTC
        22
        2307
        0.0000%
        TCTGTTCACACTGGTGTTGTCTCCATCCCTGCCAAAGCTGCCTTGCTGCC
        26
        2388194
        0.0451%
        TCTTCTGTCATCTGGGGCTTGCGCTTGACGAGGTTGACGAGCAATTTCAG
        15
        235
        0.0000%
        TGATGATTGCTGGGAATGGCTTTCCACTGCTGTCTTTCTGATAGAGAGTC
        14
        178
        0.0000%
        TTATTTCAGGTATGTAAAGGTGTCCAGAGGAGGGGATCTGTCTTTAGACT
        20
        265
        0.0000%

        Adapter Content

        The cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position.

        Note that only samples with ≥ 0.1% adapter contamination are shown.

        There may be several lines per sample, as one is shown for each adapter detected in the file.

        From the FastQC Help:

        The plot shows a cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position. Once a sequence has been seen in a read it is counted as being present right through to the end of the read so the percentages you see will only increase as the read length goes on.

        No samples found with any adapter contamination > 0.1%

        Status Checks

        Status for each FastQC section showing whether results seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        FastQC assigns a status for each section of the report. These give a quick evaluation of whether the results of the analysis seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        It is important to stress that although the analysis results appear to give a pass/fail result, these evaluations must be taken in the context of what you expect from your library. A 'normal' sample as far as FastQC is concerned is random and diverse. Some experiments may be expected to produce libraries which are biased in particular ways. You should treat the summary evaluations therefore as pointers to where you should concentrate your attention and understand why your library may not look random and diverse.

        Specific guidance on how to interpret the output of each module can be found in the relevant report section, or in the FastQC help.

        In this heatmap, we summarise all of these into a single heatmap for a quick overview. Note that not all FastQC sections have plots in MultiQC reports, but all status checks are shown in this heatmap.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Software Versions

        Software Versions lists versions of software tools extracted from file contents.

        SoftwareVersion
        FastQC0.11.9